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Mast cell-expressed orphan receptor CCRL2 binds chemerin and is required for optimal induction of IgE-mediated passive cutaneous anaphylaxis.

Zabel BA, Nakae S, Zúñiga L, Kim JY, Ohyama T, Alt C, Pan J, Suto H, Soler D, Allen SJ, Handel TM, Song CH, Galli SJ, Butcher EC - J. Exp. Med. (2008)

Bottom Line: Mast cells contribute importantly to both protective and pathological IgE-dependent immune responses.In contrast to other "silent" or professional chemokine interreceptors, chemerin binding does not trigger ligand internalization.Rather, CCRL2 is able to bind the chemoattractant and increase local concentrations of bioactive chemerin, thus providing a link between CCRL2 expression and inflammation via the cell-signaling chemerin receptor CMKLR1.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology and Vascular Biology, Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA. bazabel@alum.mit.edu

ABSTRACT
Mast cells contribute importantly to both protective and pathological IgE-dependent immune responses. We show that the mast cell-expressed orphan serpentine receptor mCCRL2 is not required for expression of IgE-mediated mast cell-dependent passive cutaneous anaphylaxis but can enhance the tissue swelling and leukocyte infiltrates associated with such reactions in mice. We further identify chemerin as a natural nonsignaling protein ligand for both human and mouse CCRL2. In contrast to other "silent" or professional chemokine interreceptors, chemerin binding does not trigger ligand internalization. Rather, CCRL2 is able to bind the chemoattractant and increase local concentrations of bioactive chemerin, thus providing a link between CCRL2 expression and inflammation via the cell-signaling chemerin receptor CMKLR1.

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Mast cell–expressed mCCRL2 is required for maximal tissue swelling and numbers of dermal leukocytes in PCA. (A) WT or CCLR2 KO mice were sensitized by injection of 50 ng of anti-DNP IgE into left ear skin (with vehicle injection into right ear skin as the control). The mice were challenged by i.v. injection of DNP-HSA (200 μg/mouse) the next day, and ear swelling was measured at the indicated time points (mean ± SEM; n = 3 experiments with a total of 21 KO and 16 WT mice per group). *, P < 0.005 by ANOVA comparing swelling in WT vs. KO ears sensitized with antigen-specific IgE. (B–D) The ears of mast cell–deficient KitW-sh/Wsh mice were engrafted with BMCMCs from either WT or mCCRL2 KO mice. 6–8 wk later, the mice were sensitized (5 ng IgE/left ear, with vehicle into the right ear as the control), challenged with specific antigen (200 μg DNP-HSA i.v.), and assessed for tissue swelling (B), as described in A, and for numbers of mast cells (C) or leukocytes (D) per millimeters squared of dermis. Data are shown as mean ± SEM, n = 3 experiments, with 15 total mice per group in B, and the numbers of mice sampled for histological data are shown in C and D. *, P < 0.001 by ANOVA comparing swelling in mCCRL2 KO BMCMC- vs. WT BMCMC-engrafted ears sensitized with antigen-specific IgE. (C) Enumeration of mast cells present in the dermis of ear skin in engrafted animals from B after elicitation of PCA (IgE) or in vehicle-injected control (vehicle) ears. **, P < 0.005 by Student's t test versus values for the vehicle-injected ears in the corresponding WT BMCMC- or KO BMCMC-engrafted KitW-sh/Wsh mice. (D) Numbers of leukocytes per millimeters squared of dermis, assessed in formalin-fixed paraffin-embedded hematoxylin and eosin–stained sections of mice from B and C. ***, P < 0.0001 by the Mann Whitney U test versus corresponding values for the vehicle-injected ears in WT BMCMC- or KO BMCMC-engrafted KitW-sh/Wsh mice. The numbers over the bars for vehicle-injected mice are the mean values.
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fig4: Mast cell–expressed mCCRL2 is required for maximal tissue swelling and numbers of dermal leukocytes in PCA. (A) WT or CCLR2 KO mice were sensitized by injection of 50 ng of anti-DNP IgE into left ear skin (with vehicle injection into right ear skin as the control). The mice were challenged by i.v. injection of DNP-HSA (200 μg/mouse) the next day, and ear swelling was measured at the indicated time points (mean ± SEM; n = 3 experiments with a total of 21 KO and 16 WT mice per group). *, P < 0.005 by ANOVA comparing swelling in WT vs. KO ears sensitized with antigen-specific IgE. (B–D) The ears of mast cell–deficient KitW-sh/Wsh mice were engrafted with BMCMCs from either WT or mCCRL2 KO mice. 6–8 wk later, the mice were sensitized (5 ng IgE/left ear, with vehicle into the right ear as the control), challenged with specific antigen (200 μg DNP-HSA i.v.), and assessed for tissue swelling (B), as described in A, and for numbers of mast cells (C) or leukocytes (D) per millimeters squared of dermis. Data are shown as mean ± SEM, n = 3 experiments, with 15 total mice per group in B, and the numbers of mice sampled for histological data are shown in C and D. *, P < 0.001 by ANOVA comparing swelling in mCCRL2 KO BMCMC- vs. WT BMCMC-engrafted ears sensitized with antigen-specific IgE. (C) Enumeration of mast cells present in the dermis of ear skin in engrafted animals from B after elicitation of PCA (IgE) or in vehicle-injected control (vehicle) ears. **, P < 0.005 by Student's t test versus values for the vehicle-injected ears in the corresponding WT BMCMC- or KO BMCMC-engrafted KitW-sh/Wsh mice. (D) Numbers of leukocytes per millimeters squared of dermis, assessed in formalin-fixed paraffin-embedded hematoxylin and eosin–stained sections of mice from B and C. ***, P < 0.0001 by the Mann Whitney U test versus corresponding values for the vehicle-injected ears in WT BMCMC- or KO BMCMC-engrafted KitW-sh/Wsh mice. The numbers over the bars for vehicle-injected mice are the mean values.

Mentions: We next examined a mast cell–dependent model of atopic allergy, the IgE-dependent PCA reaction. Animals sensitized with 150 ng/ear DNP-specific IgE and challenged with antigen (2,4-DNP-conjugated human serum albumin [DNP-HSA]) i.v. developed strong local inflammatory responses, with no significant difference in the tissue swelling observed in WT versus CCRL2 KO mice (82 ± 9 vs. 91 ± 9 × 10−2 mm of swelling at 30 min after antigen challenge, respectively; P > 0.05, Student's t test; Fig. S4 A, available at http://www.jem.org/cgi/content/full/jem.20080300/DC1). However, when the sensitizing dose of DNP-specific IgE was reduced to 50 ng/ear, the PCA reactions in CCRL2 KO mice were significantly impaired compared with those in WT mice (42.2 ± 2.8 vs. 24.9 ± 2.7 × 10−2 mm of swelling at 30 min after antigen challenge, respectively; P < 0.005, Student's t test; Fig. 4 A).


Mast cell-expressed orphan receptor CCRL2 binds chemerin and is required for optimal induction of IgE-mediated passive cutaneous anaphylaxis.

Zabel BA, Nakae S, Zúñiga L, Kim JY, Ohyama T, Alt C, Pan J, Suto H, Soler D, Allen SJ, Handel TM, Song CH, Galli SJ, Butcher EC - J. Exp. Med. (2008)

Mast cell–expressed mCCRL2 is required for maximal tissue swelling and numbers of dermal leukocytes in PCA. (A) WT or CCLR2 KO mice were sensitized by injection of 50 ng of anti-DNP IgE into left ear skin (with vehicle injection into right ear skin as the control). The mice were challenged by i.v. injection of DNP-HSA (200 μg/mouse) the next day, and ear swelling was measured at the indicated time points (mean ± SEM; n = 3 experiments with a total of 21 KO and 16 WT mice per group). *, P < 0.005 by ANOVA comparing swelling in WT vs. KO ears sensitized with antigen-specific IgE. (B–D) The ears of mast cell–deficient KitW-sh/Wsh mice were engrafted with BMCMCs from either WT or mCCRL2 KO mice. 6–8 wk later, the mice were sensitized (5 ng IgE/left ear, with vehicle into the right ear as the control), challenged with specific antigen (200 μg DNP-HSA i.v.), and assessed for tissue swelling (B), as described in A, and for numbers of mast cells (C) or leukocytes (D) per millimeters squared of dermis. Data are shown as mean ± SEM, n = 3 experiments, with 15 total mice per group in B, and the numbers of mice sampled for histological data are shown in C and D. *, P < 0.001 by ANOVA comparing swelling in mCCRL2 KO BMCMC- vs. WT BMCMC-engrafted ears sensitized with antigen-specific IgE. (C) Enumeration of mast cells present in the dermis of ear skin in engrafted animals from B after elicitation of PCA (IgE) or in vehicle-injected control (vehicle) ears. **, P < 0.005 by Student's t test versus values for the vehicle-injected ears in the corresponding WT BMCMC- or KO BMCMC-engrafted KitW-sh/Wsh mice. (D) Numbers of leukocytes per millimeters squared of dermis, assessed in formalin-fixed paraffin-embedded hematoxylin and eosin–stained sections of mice from B and C. ***, P < 0.0001 by the Mann Whitney U test versus corresponding values for the vehicle-injected ears in WT BMCMC- or KO BMCMC-engrafted KitW-sh/Wsh mice. The numbers over the bars for vehicle-injected mice are the mean values.
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fig4: Mast cell–expressed mCCRL2 is required for maximal tissue swelling and numbers of dermal leukocytes in PCA. (A) WT or CCLR2 KO mice were sensitized by injection of 50 ng of anti-DNP IgE into left ear skin (with vehicle injection into right ear skin as the control). The mice were challenged by i.v. injection of DNP-HSA (200 μg/mouse) the next day, and ear swelling was measured at the indicated time points (mean ± SEM; n = 3 experiments with a total of 21 KO and 16 WT mice per group). *, P < 0.005 by ANOVA comparing swelling in WT vs. KO ears sensitized with antigen-specific IgE. (B–D) The ears of mast cell–deficient KitW-sh/Wsh mice were engrafted with BMCMCs from either WT or mCCRL2 KO mice. 6–8 wk later, the mice were sensitized (5 ng IgE/left ear, with vehicle into the right ear as the control), challenged with specific antigen (200 μg DNP-HSA i.v.), and assessed for tissue swelling (B), as described in A, and for numbers of mast cells (C) or leukocytes (D) per millimeters squared of dermis. Data are shown as mean ± SEM, n = 3 experiments, with 15 total mice per group in B, and the numbers of mice sampled for histological data are shown in C and D. *, P < 0.001 by ANOVA comparing swelling in mCCRL2 KO BMCMC- vs. WT BMCMC-engrafted ears sensitized with antigen-specific IgE. (C) Enumeration of mast cells present in the dermis of ear skin in engrafted animals from B after elicitation of PCA (IgE) or in vehicle-injected control (vehicle) ears. **, P < 0.005 by Student's t test versus values for the vehicle-injected ears in the corresponding WT BMCMC- or KO BMCMC-engrafted KitW-sh/Wsh mice. (D) Numbers of leukocytes per millimeters squared of dermis, assessed in formalin-fixed paraffin-embedded hematoxylin and eosin–stained sections of mice from B and C. ***, P < 0.0001 by the Mann Whitney U test versus corresponding values for the vehicle-injected ears in WT BMCMC- or KO BMCMC-engrafted KitW-sh/Wsh mice. The numbers over the bars for vehicle-injected mice are the mean values.
Mentions: We next examined a mast cell–dependent model of atopic allergy, the IgE-dependent PCA reaction. Animals sensitized with 150 ng/ear DNP-specific IgE and challenged with antigen (2,4-DNP-conjugated human serum albumin [DNP-HSA]) i.v. developed strong local inflammatory responses, with no significant difference in the tissue swelling observed in WT versus CCRL2 KO mice (82 ± 9 vs. 91 ± 9 × 10−2 mm of swelling at 30 min after antigen challenge, respectively; P > 0.05, Student's t test; Fig. S4 A, available at http://www.jem.org/cgi/content/full/jem.20080300/DC1). However, when the sensitizing dose of DNP-specific IgE was reduced to 50 ng/ear, the PCA reactions in CCRL2 KO mice were significantly impaired compared with those in WT mice (42.2 ± 2.8 vs. 24.9 ± 2.7 × 10−2 mm of swelling at 30 min after antigen challenge, respectively; P < 0.005, Student's t test; Fig. 4 A).

Bottom Line: Mast cells contribute importantly to both protective and pathological IgE-dependent immune responses.In contrast to other "silent" or professional chemokine interreceptors, chemerin binding does not trigger ligand internalization.Rather, CCRL2 is able to bind the chemoattractant and increase local concentrations of bioactive chemerin, thus providing a link between CCRL2 expression and inflammation via the cell-signaling chemerin receptor CMKLR1.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology and Vascular Biology, Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA. bazabel@alum.mit.edu

ABSTRACT
Mast cells contribute importantly to both protective and pathological IgE-dependent immune responses. We show that the mast cell-expressed orphan serpentine receptor mCCRL2 is not required for expression of IgE-mediated mast cell-dependent passive cutaneous anaphylaxis but can enhance the tissue swelling and leukocyte infiltrates associated with such reactions in mice. We further identify chemerin as a natural nonsignaling protein ligand for both human and mouse CCRL2. In contrast to other "silent" or professional chemokine interreceptors, chemerin binding does not trigger ligand internalization. Rather, CCRL2 is able to bind the chemoattractant and increase local concentrations of bioactive chemerin, thus providing a link between CCRL2 expression and inflammation via the cell-signaling chemerin receptor CMKLR1.

Show MeSH
Related in: MedlinePlus